In Planta Delivery of Chemical Compounds into Barley Meiocytes: EdU as Compound Example
Here, we describe a protocol for in planta delivery of chemical compounds into meiocytes of different barley genotypes not impacting plant fertility allowing to harvest seeds from treated plants. Compound uptake into meiocytes is assessed by determining 5-ethynyl-2′-deoxyuridine (EdU) incorporation. Similar to EdU, other compounds being soluble in an aqueous solution can be delivered in planta before/during meiosis to decipher their impact on meiosis and meiotic recombination.
We give practical advice on how to deliver EdU as compound example (delivery via injection or needle and thread, addition of detergents or surfactants to increase compound uptake), how in planta compound delivery can be established for your plant material under specific growing conditions, how to generate and characterize barley hybrid plants, and how to conduct a meiotic cytological study of (treated) barley plants.
Key wordsMeiotic recombination In planta compound delivery Meiosis EdU (5-ethynyl-2′-deoxyuridine) Cytology Hordeum vulgare Kompetitive allele specific PCR (KASP) analysis
We would like to thank the gardeners team from the IPK Gatersleben for their excellent support with plant cultivation, Jana Lorenz for excellent technical assistance, all members of the Meiosis team at the IPK for fruitful discussions, as well as the German Federal Ministry of Education and Research (BMBF—031B0188), Bayer CropScience AG (Grants4Traits 2016-1-18), the Marie-Curie “MEICOM” network H2020 ITN-765212, and the IPK Gatersleben for financial support.
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